Int. J. Mol. Sci. 2018, 19(1), 85; https://doi.org/10.3390/ijms19010085
In Vitro Selection of a Single-Stranded DNA Molecular Recognition Element against the Pesticide Fipronil and Sensitive Detection in River Water
Ka L. Hong 1,*
and Letha J. Sooter 2
and Letha J. Sooter 21
Department of Pharmaceutical Sciences, Nesbitt School of Pharmacy, Wilkes University, 84 W. South Street, Wilkes-Barre, PA 18766, USA
2
Independent Researcher, Pittsburgh, PA 15234, USA
*
Author to whom correspondence should be addressed.
Received: 31 October 2017 / Revised: 12 December 2017 / Accepted: 25 December 2017 / Published: 28 December 2017
(This article belongs to the Special Issue Aptamers)
Abstract
Fipronil is a commonly used insecticide that has been shown to have environmental and human health risks. The current standard methods of detection for fipronil and its metabolites, such as GC-MS, are time consuming and labor intensive. In this study, a variant of systematic evolution of ligands by exponential enrichment (SELEX), was utilized to identify the first single-stranded DNA (ssDNA) molecular recognition element (MRE) that binds to fipronil with high affinity (Kd = 48 ± 8 nM). The selected MRE displayed low cross binding activity on various environmentally relevant, structurally unrelated herbicides and pesticides, in addition to broad-spectrum binding activity on major metabolites of fipronil and a structurally similar pesticide in prepared river samples. Additionally, a proof-of-principle fluorescent detection assay was developed by using the selected ssDNA MRE as a signal-reporting element, with a limit of detection of 105 nM in a prepared river water sample. View Full-TextKeywords:
SELEX; in vitro selection; aptamer; molecular recognition element (MRE); fipronil; pesticide; insecticide; ELISA; Decoy-SELEX
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Hong, K.L.; Sooter, L.J. In Vitro Selection of a Single-Stranded DNA Molecular Recognition Element against the Pesticide Fipronil and Sensitive Detection in River Water. Int. J. Mol. Sci. 2018, 19, 85.
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